Phenolic composition by UHPLC-Q-TOF-MS/MS and stability of anthocyanins from Clitoria ternatea L. (butterfly pea) blue petals.

The aim of the present study was to evaluate the phenolic composition of crude lyophilized extracts (CLE) and partially purified (PPE) extracts of C. ternatea blue petals as well as the anthocyanin stability against pH, temperature and light in the presence and absence of fructooligosaccharides. Twelve compounds were tentatively identified by UHPLC-Q-TOF-MS/MS in CLE and PPE extracts. In direct/reverse spectrophotometric titration, anthocyanins showed colour changes between pH 2.25 to 10.20, and colour reversibility, maintaining antioxidant activity against the DPPH radical. The aqueous extracts at pH 3.6 and 5.4 exhibited thermal stability with the presence and absence of fructooligosaccharides with activation energy higher than 99 kJ/mol. The addition of fructooligosaccharides in the extracts at pH 5.4 exposed to light provided a protective effect against anthocyanin photodegradation. The data show the technological potential of aqueous extract of C. ternatea blue petals as a natural colourant in a functional beverage model system.

Response surface optimization of phenolic compounds from jabuticaba (Myrciaria cauliflora [Mart.] O.Berg) seeds: Antioxidant, antimicrobial, antihyperglycemic, antihypertensive and cytotoxic assessments.

The aim of this study was to evaluate the effects of different solvents and maximize the extraction of bioactive compounds from jabuticaba (Myrciaria cauliflora) seeds. In general, the solvent system composed of water and propanone (52:48 v/v) modified the extract polarity and increased extraction yield of bioactive compounds. The optimized extract presented antioxidant capacity measured by different chemical and biological assays. The optimized extract exerted antiproliferative and cytotoxic effects against A549 and HCT8 cells, antimicrobial and antihemolytic effects, inhibited α-amylase/α-glucosidase activities and presented in vitro antihypertensive effect. Nonetheless, the optimized extract showed no cytotoxicity in a human cell model (IMR90). Vescalagin, castalagin and ellagic acid were the major phenolic compounds in the optimized extract. Our results show that jabuticaba seed may be a potential ingredient for the development of potentially functional foods.

Clitoria ternatea L. petal bioactive compounds display antioxidant, antihemolytic and antihypertensive effects, inhibit α-amylase and α-glucosidase activities and reduce human LDL cholesterol and DNA induced oxidation.

The purpose of this study was to use a statistical approach to optimise the experimental conditions regarding the extraction of bioactive compounds, and to analyse the in vitro functional properties of crude lyophilized extracts (CLE) and partially purified (PPE) extracts of Clitoria ternatea petals. The results showed that the factors of temperature and time influenced the extraction of phenolic compounds, antioxidant activity and the physicochemical parameters. Simultaneous optimisation showed that the same levels of bioactive compounds were extracted when using temperatures from 11.7 to 68.3 °C and times from 8.47 to 51.12 min. Principal component analysis revealed the experimental conditions that provided the extraction producing the highest level of phenolic content (40 °C/30 min). The CLE showed antimicrobial activity; protective effect against hemolysis of erythrocytes; inhibition of α-amylase, α-glucosidase and angiotensin-I-converting (ACE-I) enzymes; and inhibition of lipid peroxidation. The CLE and PPE demonstrated oxygen radical absorption capacity; inhibition of DNA strand scission; inhibition of LDL cholesterol oxidation; intracellular antioxidant activity against reactive oxygen species (>100 µg/mL); and no cytotoxicity (IC50, GI50 and LC50 > 900 µg/mL) against A549, HCT8 and IMR90 cell lines.

Chemometric Authentication of Brazilian Coffees Based on Chemical Profiling.

In this work, different chemometric tools were compared to classify n = 26 conventional (CONV) and n = 19 organic (ORG) coffees from the main Brazilian producing regions based on the chemical composition, physicochemical properties, and antioxidant activity. Principal component analysis separated ORG and CONV coffees but the distinction among the producing regions of Brazilian coffee was not possible. Partial least squares discriminant analysis classified all ORG and CONV coffees in the external validation. Similarly, linear discriminant analysis was able to discriminate 100% and 81% of ORG and CONV coffees in the external validation, respectively, in which total phenolic content (TPC), ferric reducing antioxidant activity, and caffeic acid were the main discriminant variables. Overall 100% of samples from Paraná, Minas Gerais, and blended samples were correctly classified, where TPC, flavonoids, inhibition of lipid peroxidation, caffeic acid, pH, and soluble solids were the main discriminant variables. Support vector machines classified 95% ORG and 88% CONV, 100% Coffea arabica, and 88% and 78% coffees produced in São Paulo and Minas Gerais. k-Nearest neighbors was effective in distinguishing 100% CONV, 89% ORG, 100% coffees from São Paulo, and 100% C. arabica coffees. Overall, HPLC data and simple physicochemical parameters allied to chemometrics were effective in authenticating the cultivation system and the botanical origin of Brazilian coffees. PRACTICAL APPLICATION: Coffee adulteration is a serious problem in the food chain as some fraudsters replace coffee powder by other cheaper products. In the case of organic coffee, this scenario is even worse as still there is not a universal method to differentiate conventionally grown coffee from its organic counterpart. In addition, Brazilian coffee is produced in different regions and the commercial value varies. Therefore, we analyzed some physicochemical, chemical, and antioxidant properties of Brazilian coffees from distinct origins and classified the samples using chemometrics. Our approach seems to be interesting for quality control purposes.

Red Chicory (Cichorium intybus) Extract Rich in Anthocyanins: Chemical Stability, Antioxidant Activity, and Antiproliferative Activity In Vitro.

Red chicory leaves are appreciated sensorially and their constituents contain bioactive properties. The objectives of this study were as follows: to use an experimental design to extract anthocyanins from red chicory in aqueous solution at pH 2.5; to determine the stability of the extracts in relation to temperature and pH; and to evaluate the antioxidant activity and in vitro cytotoxic effect of the lyophilized and purified extracts. The best extraction conditions for the bioactive compounds from red chicory were a temperature of 64.2 °C for 25 min; the anthocyanin content was 73.53 ± 0.13 mg per 100 g fresh weight basis sample. The EC50 (Half maximal effective concentration) value for the antioxidant activity assay in relation to DPPH (2,2-diphenyl-1-picrylhydrazyl) with optimized extract was 0.363, which corresponds to a concentration of 39.171 µmol/L of anthocyanins. The activation energy for the degradation reaction of the anthocyanins from the red chicory extract was 84.88 kJ/mol. The optimized extract, which was rich in anthocyanins, showed chemical and biological antioxidant activity (protection against erythrocyte hemolysis) and inhibited lipid peroxidation in vitro. The Cichorium intybus L. extracts interfered on the levels of reactive oxygen species generation and the crude extract did not present procarcinogenic effect. PRACTICAL APPLICATION: Red chicory is basically consumed as a part of traditional dishes worldwide. Here, we developed a process to extract and purify the anthocyanins from Cichorium intybus leaves and test the extracts in terms of the chemical composition, thermal stability, antioxidant activity, and antiproliferative effects. The anthocyanin-rich extract presented antioxidant activity in chemical and biological assays and low cytotoxicity and cytoprotective effects in relation to HepG2, HCT8, and Caco-2 cell lines. Additionally, the red chicory extract protected human erythrocytes against hemolysis. This extract may be used as a natural colorant/antioxidant in foods.

Chemical study, antioxidant, anti-hypertensive, and cytotoxic/cytoprotective activities of Centaurea cyanus L. petals aqueous extract.

This study aimed to optimise the experimental conditions of extraction of the phytochemical compounds and functional properties of Centaurea cyanus petals. The following parameters were determined: the chemical composition (LC-ESI-MS/MS), the effects of pH on the stability and antioxidant activity of anthocyanins, the inhibition of lipid peroxidation, antioxidant activity, anti-hemolytic activity, antimicrobial, anti-hypertensive, and cytotoxic/cytoprotective effect, and the measurements of intracellular reactive oxygen species. Results showed that the temperature and time influenced (p ≤ 0.05) the content of flavonoids, anthocyanins, and FRAP. Only the temperature influenced the total phenolic content, non-anthocyanin flavonoids, and antioxidant activity (DPPH). The statistical approach made it possible to obtain the optimised experimental extraction conditions to increase the level of bioactive compounds. Chlorogenic, caffeic, ferulic, and p-coumaric acids, isoquercitrin, and coumarin were identified as the major compounds in the optimised extract. The optimised extract presented anti-hemolytic and anti-hypertensive activity in vitro, in addition to showing stability and reversibility of anthocyanins and antioxidant activity with pH variation. The C. cyanus petals aqueous extract exhibited high IC50 and GI50 (>900 µg/mL) values for all cell lines, meaning low cytotoxicity. Based on the stress oxidative assay, the extract exhibited pro-oxidant action (10-100 µg/mL) but did not cause damage or cell death.

Hibiscus sabdariffa anthocyanins-rich extract: Chemical stability, in vitro antioxidant and antiproliferative activities.

Hibiscus sabdariffa calyx is a rich source of anthocyanins and other bioactive compounds but no study reported the effects of experimental conditions on the extraction of these chemical compounds. Therefore, the effects of time and extraction temperature on the bioactive compounds and antioxidant activity of Hibiscus sabdariffa calyx were evaluated. In addition, the effects of copigmentation and pH on the stability of anthocyanins were assessed and the cytotoxic effects (LC50, IC50, and GC50) of the extracts were determined in relation to tumor cell lines - Caco-2, HepG-2, HCT8, and A549. The temperature significantly influenced the total anthocyanins and flavonoids contents. The interaction between time/temperature influenced the total phenolic content and ascorbic acid. The t1/2 and the percentage of colour retention decreased markedly at temperatures above 80 °C. Variations in pH conserved the antioxidant activity of the anthocyanins, and the protonation-deprotonation process of the extract was reversible. The treatment of cells with purified anthocyanin extract or crude extracts at 5-800 µg mL-1 did not show significant cytotoxic effects on the cell lines, corroborating the chemical antioxidant effect of the extracts (DPPH assay). Cyanidin-3-glucoside, delphinidin-3-sambubioside, delphinidin-3-glucoside, and cyanidin-3-sambubioside were identified in the extracts by LC-ESI-MS.

Jabuticaba (Myrciaria cauliflora) Seeds: Chemical Characterization and Extraction of Antioxidant and Antimicrobial Compounds.

This study was aimed to assess the effect of time and temperature on the extraction of antioxidant compounds from jabuticaba seeds (Myrciaria cauliflora cv. Sabará), to optimize the solvent proportion (water, ethyl alcohol, and propanone), and to characterize the extract according to the chemical composition, antioxidant, and antimicrobial properties. Proximal composition, total phenolic content (TPC), antioxidant, and antimicrobial activities were analyzed. The optimized solvent ratio of 60% water and 40% propanone provided a mean TPC of 8.65 g GAE/100 g seeds and the antioxidant activity toward 2,2-diphenyl-1-picrylhydrazyl (DPPH) was 82.79% ± 0.50%. Time and temperature parameters did not influence the yield of TPC. The gross seed extract was partially purified and both exhibited a high antioxidant activity and antimicrobial potential toward Gram-positive and Gram-negative bacteria. The purified jabuticaba seed lyophilized extract contained a higher (P < 0.05) TPC, o-diphenols, flavonols, and antioxidant activity measured by the DPPH assay and total reducing capacity as compared to the gross lyophilized extract. Electrospray ionization coupled with tandem mass spectrometry (ESI-MS/MS) data showed the presence of ellagitannins and ellagic acid in the extracts, which are probably the responsible for the antimicrobial and antioxidant activities.

Comparison between Folin-Ciocalteu and Prussian Blue Assays to Estimate The Total Phenolic Content of Juices and Teas Using 96-Well Microplates.

UNLABELLED: Folin-Ciocalteu colorimetric assay (FC) is the most widely used assay to estimate the total phenolic content in foods, beverages, herbs and other plant extracts, but many chemical compounds may act as interfering agents, producing inaccurate estimations of the real concentration of phenolic compounds in the matrix. Based on this limitation, the objective of this study was to compare, quantitatively, the Folin-Ciocalteu and Prussian Blue (PB) assays in estimating the total phenolic content in purple grape juices (n = 20; Vitis labrusca L.) and teas (n = 25) from different botanical origins using 96-well microplates. PB assay presented a low limit of detection (PB = 0.27 mg/L; FC = 0.25 mg/L) and quantification (PB = 0.92 mg/L; FC = 0.82 mg/L), showing its suitability in screening the total phenolic content in grape juices and teas. FC and PB assays presented a high association (P < 0.0001) for teas (r = 0.887) and grape juices (r = 0.923). The advantages of PB over FC assay are its simplicity, low time consumption (15 min reaction as compared to 60 min reaction for the FC assay), lower usage of reagents (solutions are prepared in a mM base), and higher selectivity. Additionally, PB assay was proven to be reproducible and repeatable and, therefore, may be used as an alternative to FC assay. PRACTICAL APPLICATION: Prussian Blue assay (PB) has been used as an alternative to Folin-Ciocalteu assay (FC) to estimate the total content of phenolic compounds in herbs and some natural products. In our study we showed that the advantages of PB assay over FC are its simplicity, low time consumption (15 min reaction as compared to 60 min reaction for the FC assay), lower usage of reagents (solutions are prepared in a mM base) and higher selectivity as compared to FC assay. Additionally, PB assay was proven to be reproducible and repeatable and, therefore, may be used as an alternative to FC assay.

Cytotoxic effect of inositol hexaphosphate and its Ni(II) complex on human acute leukemia Jurkat T cells.

Inositol hexaphosphate (InsP6) is present in cereals, legumes, nuts and seed oils and is biologically active against some tumor and cancer cells. Herein, this study aimed at evaluating the cellular toxicity, antiproliferative activity and effects on cell cycle progression of free InsP6 and InsP6-Ni(II) of leukemic T (Jurkat) and normal human cells. Treatments with InsP6 at concentrations between 1.0 and 4.0mM significantly decreased the viability of Jurkat cells, but showed no cytotoxic effect on normal human lymphocytes. Treatment with InsP6-Ni(II) complex at concentrations between 0.05 and 0.30 mM showed an anti-proliferative dose and a time-dependent effect, with significantly reduced cell viability of Jurkat cells but showed no cytotoxic effect on normal human lymphocytes as compared to the control. Ni(II) free ion was toxic to normal cells while InsP6-Ni(II) had no cytotoxic effect. The InsP6-Ni(II) complex potentiated (up to 10×) the antiproliferative effect of free InsP6 on Jurkat cells. The cytometric flow assay showed that InsP6 led to an accumulation of cells in the G0/G1 phase of the cell cycle, accompanied by a decrease in the number of cells in S and G2/M phases, whereas InsP6-Ni(II) has led to an accumulation of cells in the S and G2/M phases. Our findings showed that InsP6-Ni(II) potentiates cytotoxic effects of InsP6 on Jurkat cells and may be a potential adjuvant in the treatment of cancer.